1. Field of the Invention
The invention disclosed herein relates generally to mixing of fluid samples using shakers for microplates, small diameter test tubes, and like-configured fluid containers, and more particularly to mixing of fluid samples using a multidirectional shaker of simplified construction comprising a support tray resiliently mounted above a base through a plurality of spring members arranged in differing directions, and a plurality of electromagnetic drives or mechanical drives for imparting at least bi-directional vibratory motion to the support tray in order to mix the contents of a microplate or collection of specimen tubes positioned on the support tray, irrespective of the diameter of the microplate wells or tubes.
2. Description of the Background
The processing of biological specimens or chemical products in laboratories often requires the mixing of analytes within a container in order to carry out a desired reaction. Such containers have often comprised beakers or flasks whose contents were traditionally mixed by either manually shaking the beaker or flask, or by using a stirring rod. Other mixing apparatus have included a Teflon coated magnet placed within a beaker or flask and driven magnetically in a rotary motion to mix the beaker or flask contents. Unfortunately, manually shaking the beaker or flask provides insufficient means to control the mixing of the contents and easily results in laboratory technicians accidentally dropping the container and ruining the sample. Likewise, the use of stirring rods has required that the laboratory technician either thoroughly wash the rod between specimens in order to avoid cross-contamination, or throw away and replace disposable rods for applications with large numbers of specimens, making the rapid mixing of large numbers of specimens highly impractical.
In order to overcome these shortcomings, motor driven orbital shakers were developed which enabled a laboratory technician to place a beaker or flask on a motor driven platform that would cause the beaker or flask to travel in a continuous orbit to mix its contents. So long as the diameter of the beaker or flask holding a sample is greater than the orbit diameter of the platform, mixing of the contents will occur. For example, as shown in the schematic view of a prior art orbital mixer of FIG. 1a, the center of the flask travels in an orbital path equivalent to the orbit of the platform, and the centrifugal forces on the liquid will reverse every 180° to provide adequate mixing of the contents.
However, as the number of specimens needed to be analyzed in a given time period has grown, the quest for efficiency in the processing of such specimens has resulted in smaller and smaller sample sizes being studied, and thus smaller and smaller containers for holding those samples. Unfortunately, as smaller sized beakers and flasks were used, those orbital shakers having an orbit diameter that was larger than the beaker or flask diameter were shown to be ineffective for mixing the contents. For example, as shown in the schematic view of a prior art orbital mixer of FIG. 1b, a beaker or flask having a diameter that is smaller than the orbit diameter of the mixer simply travels in the shaker's orbit, and centrifugal forces drive the liquid contained within the beaker or flask against the side of the container which is furthest from the center of orbit. If there are any suspended solids in the liquid, they will likewise be driven against the outside wall of the container, and fail to mix with the solution. In order to alleviate this problem, a few orbital shakers have been made available having orbit diameters of as little as ⅛″.
As the need for processing greater numbers of samples in shorter amounts of time continued to grow, microplates were developed to hold multiple samples of a chemical or biological material to be analyzed in a single, compact structure having a rectangular grid of a large number of distinct “wells.” Such microplates are available today in 96-well, 384-well, and even 1536-well configurations. Likewise, racks of small diameter tubes have been developed providing a similar array of specimen-holding chambers. Obviously, the greater the number of wells or tubes in a standard microplate footprint, the smaller the diameter of the well, such that for microplates and tubes having chamber diameters of far less than ⅛″, an orbit of far less than ⅛″ would likewise be required in order to ensure proper mixing. As was true with orbital mixers for large flasks, the contents of such a small diameter tube rotating in an orbit larger than its own diameter are difficult to mix. Using an orbit larger than the well or tube diameter causes the liquid contents to move to the outside of the orbit and rise up the inner wall of the tube which is closest to the outside radius of the orbit. The contents of the tube begin to spin inside the tube with a relatively small amount of relative motion (or shearing) between adjacent layers of fluid within the walls of the tube. As the orbital speed is increased, the liquid in the tube is forced outward by centrifugal force, rising up the inner wall of the tube until it spills over the top. Given the orbit diameter limitation of only ⅛″, traditional horizontal orbital shakers have thus been ineffective in shaking microplates and tube collections having such small diameter chambers.
Given the failure of traditional orbiting mixing apparatus to provide an effective means of mixing the contents of small well microplates and small diameter tubes, attempts have been made to provide mixing apparatus specifically configured for mixing the contents of microplate wells, but unfortunately have also met with little success. For example, U.S. Pat. No. 3,635,446 to Kurosawa et al. discloses a microplate shaking device using an eccentric motor to uncontrollably vibrate a microplate holding plate through a horizontal plane. Likewise, U.S. Pat. No. 4,102,649 to Sasaki discloses a microplate shaker device which pivotally mounts a microplate to a vibration plate, and slidably mounts the microplate atop a number of props. The vibration plate is caused to vibrate by either an electromagnet or an eccentric wheel in a nonlinear, horizontal manner. Further, U.S. Pat. No. 4,264,559 to Price discloses a mixing device for a specimen holder comprising two springlike metal rods upon which a specimen holder is mounted, the rods being fixed at one end in a vertical block, and a weight positioned adjacent the opposite end of the rods. Manually plucking one of the rods imparts a “pendulum-like” vibration to both rods, and thus to the specimen holder. Finally, U.S. Pat. No. 5,921,477 to Tomes et al. discloses an agitating apparatus for a “well plate holder” which comprises a vertically-oriented reciprocating saw as a means for vertically shaking a multi-well plate, and provides agitating members comprising small diameter copper or stainless steel balls within each well.
Unfortunately, none of the known prior art devices have been able to provide controlled, multidirectional vibration to a microplate or collection of small diameter tubes in order to create vibratory motion of sufficient turbulence to thoroughly mix the well or tube contents.
Furthermore, U.S. Pat. No. 5,427,451 to Schmidt discloses a mixer which utilizes a complex, microprocessor-controlled circuit to provide oscillatory drives comprised of permanent magnets and drive coils juxtaposed therewith, with each coil being independently energized by separate variable frequency sources. The drive circuits are configured to alternately attract and repel the permanent magnets so as to provide the oscillatory motion, thus requiring actuation of the drive coils at all times during operation of the mixer. Such a construction is highly complex, requiring precise control of the timing of each drive cycle, and exhibits high energy requirements for its operation. It would be highly advantageous to provide a simplified mixing construction that has a lower energy requirement, but that can still provide consistent, reliable mixing through controlled multidirectional shaking of test specimen containers.
Moreover, effective mixing requires that the layers of fluid within the tube vigorously move relative to each other. Simply driving the tube with a small orbital motion simply rotates the fluid within the tube as a large slug, with the only appreciable relative motion occurring between the tube wall surface and the outermost fluid layer. However, suddenly stopping the orbiting motion will cause the fluid which was driven up the outer tube wall to collapse, causing greater turbulence and thus better mixing. In fact, the rapid on and off cycling of such motion causes the creation of turbulence within the tube which can greatly facilitate the mixing of layers of fluid within the tube. While mechanically driven orbiting mixers have been previously known which attempt to provide such impulse-driven mixing, such devices have not met with commercial success. For example, mechanically driven orbiting mixers have been known which are provided a timer in the motor circuit to periodically stop the unit and then start it again. Such starting and stopping of the drive mechanism is costly, creates much wear and tear on the equipment, and most importantly, is limited as to the speed with which such a device can cycle on and off due to inertia and the ability of a motor to quickly accelerate.
It would therefore be advantageous to provide an electromagnetic, multidirectional shaker of simplified construction which will ensure the efficient mixing of the contents of microplates and small diameter tubes.